Volume 93, Issue 10, Pages (November 2007)

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Volume 93, Issue 10, Pages 3504-3514 (November 2007) Triton X-100 Partitioning into Sphingomyelin Bilayers at Subsolubilizing Detergent Concentrations: Effect of Lipid Phase and a Comparison with Dipalmitoylphosphatidylcholine Cristina Arnulphi, Jesús Sot, Marcos García-Pacios, José-Luis R. Arrondo, Alicia Alonso, Félix M. Goñi Biophysical Journal Volume 93, Issue 10, Pages 3504-3514 (November 2007) DOI: 10.1529/biophysj.107.104463 Copyright © 2007 The Biophysical Society Terms and Conditions

Figure 1 DSC thermograms corresponding to the gel-fluid transition of phospholipid vesicles incubated with increasing amounts of Triton. (A) SM bilayers. (B) DPPC bilayers. When appropriate, lipid/detergent mol ratios are indicated for each thermogram. Third heating scan. Biophysical Journal 2007 93, 3504-3514DOI: (10.1529/biophysj.107.104463) Copyright © 2007 The Biophysical Society Terms and Conditions

Figure 2 Temperature variation of the GP of the fluorescent probe Laurdan embedded in phospholipid bilayers. (A) SM bilayers. (B) DPPC bilayers. (●) Pure phospholipid. (○) Phospholipid/Triton (20:1mol ratio). Average values of three independent experiments±SE. The standard errors are smaller than the size of the symbols. Biophysical Journal 2007 93, 3504-3514DOI: (10.1529/biophysj.107.104463) Copyright © 2007 The Biophysical Society Terms and Conditions

Figure 3 Comparison of representative ITC isotherms for the interaction of Triton with SM at different lipid phase states: (○) gel phase (10°C), (●) fluid phase (45°C), and (□) within the lipid phase transition (38°C). Biophysical Journal 2007 93, 3504-3514DOI: (10.1529/biophysj.107.104463) Copyright © 2007 The Biophysical Society Terms and Conditions

Figure 4 Thermodynamic parameters for the interaction of Triton with SM vesicles studied as a function of temperature. (A) Partition constant, K. (B) Partition enthalpy ΔH. (C) Free energy and entropy change. Gel (g) and fluid (f) phases are indicated in the appropriate temperature ranges. Average values if three independent experiments±SE. The standard errors are often smaller than the size of the symbols. Biophysical Journal 2007 93, 3504-3514DOI: (10.1529/biophysj.107.104463) Copyright © 2007 The Biophysical Society Terms and Conditions

Figure 5 Thermodynamic parameters of the partition of Triton into DPPC membranes as a function of temperature. (A) Partition constant, K. (B) Partition enthalpy, ΔH. (C) Entropy and free energy change. Gel (g) and fluid (f) phases are indicated in the appropriate temperature ranges. Average values of three independent experiments±SE. The standard errors are sometimes smaller than the size of the symbols. Biophysical Journal 2007 93, 3504-3514DOI: (10.1529/biophysj.107.104463) Copyright © 2007 The Biophysical Society Terms and Conditions

Figure 6 Comparative values of TΔS and ΔG for the interaction of Triton with SM and DPPC membranes. (■) SM. (●) DPPC. (○) pSM. Average values of three independent experiments. SM and DPPC data are redrawn from Figs. 4 and 5, except that error bars have been omitted for the sake of clarity. For pSM data, the standard error is smaller than the size of the symbols. Biophysical Journal 2007 93, 3504-3514DOI: (10.1529/biophysj.107.104463) Copyright © 2007 The Biophysical Society Terms and Conditions